Immunoassay for equine protozoal myeloencephalitis in horses

Abstract

An immunoassay for Sarcocystis neurons antibodies in equines is described. The immunoassay uses blocking of Sarcocystis antigens by antibodies to Sarcocystis sp. other than Sarcocystis neurona in connection with the immunoassay.

We claim:  
     
       1. A kit for the detection of disease caused by  Sarcocystis neurona  which comprises in separate containers: 
       (a) an identifying antibody which specifically binds to a 30 kDa protein of  Sarcocystis neurona  as determined by SDS-gel electrophoresis under denaturing conditions; and  
       (b) a non-labeled antibody from a mammal other than an equine which is specific for a protein of a Sarcocystis sp. other than  Sarcocystis neurona.    
     
     
       2. The kit of  claim 1  which in addition contains in a further separate container the 30 kDa protein of  Sarcocystis neurona  which birds to antibodies from an equine suspected of harboring the  Sarcocystis neurona.    
     
     
       3. A kit for the detection of disease caused by  Sarcocystis neurona  in equines which comprises: 
       (a) a substrate with at least one identifying antigen of the  Sarcocystis neurona  selected from the group consisting of a 16 kDa antigen and a 30 kDa antigen when separates by SDS-PAGE electrophoresis under denaturing conditions, bound on a surface of the substrate;  
       (b) antibody to a Sarcocystis sp. other than  Sarcocystis neurona  wherein the antibody is from a mammal other than the equine; and  
       (c) at least one reagent for detection of an antibody in a fluid of the equine which antibody binds to the antigen of the  Sarcocystis neurona.    
     
     
       4. The kit of  claim 3  wherein the reagent is a labeled anti-equine antibody. 
     
     
       5. The kit of  claim 4  wherein the label is biotin which is reacted with a biotin detecting reactant. 
     
     
       6. The kit of  claim 3  wherein the substrate is used in a method selected from the group consisting of a Western blot and an ELISA. 
     
     
       7. The kit of  claim 3  wherein the substrate is a gel. 
     
     
       8. The kit of  claim 3  with a Western blot membrane as the substrate. 
     
     
       9. The kit of  claim 3  including a biotin labeled goat anti-horse IgG′ (H+L), peroxidase conjugate and aminoethyl carbazole which together provide the reagent. 
     
     
       10. The kit of  claim 1  which in addition contains in a further separate container an identifying antibody which specifically binds to a 16 kDa protein of  Sarcocystis neurona  as determined by SDS-gel electrophoresis under denaturing conditions. 
     
     
       11. The Kit of  claim 10  wherein addition contains in a further separate container the 16 kDa protein of  Sarcocystis neurona  which binds to antibodies from an equine suspected of harboring the  Sarcocystis neurona.    
     
     
       12. The kit of any one of  claim 1 ,  2 ,  10 , or  11  wherein a user of the kit is instructed to first bind proteins of  Sarcocystis neurona  with the non-labeled antibody in the kit which is from a mammal other than an equine which is specific for a protein of a Sarcocystis sp. other than  Sarcocystis neurona.    
     
     
       13. The kit of  claim 3  wherein a user of the kit is instructed to first bind antigens of  Sarcocystis neurona  with the antibody in the kit which is from a Sarcocystis sp. other than  Sarcocystis neurona  and which is from a mammal other than the equine.